Raman Imaging andScanning Electron Microscopy

RISE Microscopy®

Correlative Molecular and Ultrastructural Imaging

https://raman.oxinst.com

RISE Microscopy®

Molecular and UltrastructuralImaging
Correlative Raman imaging and scanning electron
microscopy for comprehensive sample analysis
offers a new dimension in imaging: see both theform and substance of your samples at the highest
resolution.

RISE microscopy will benefit ...
®

... researchers looking for a deeper understandingof their samples through quick and straightforward
measurements controlled with an intuitive user interface.

RISE microscopy is well suited to ...
®

... investigations in materials science, nanotechnology,forensics, geosciences, life sciences, pharmaceuticalresearch and many other fields of application.

Applications
BanataserutileCIntensity (a.u.)aRaman shift (cm-1)

Titanium dioxide(A) Two modifications of titanium dioxide, anatase and rutile, were mixed and imaged with an SEM. (B) In the Raman spectrum
anatase (blue) can be easily distinguished from rutile (red). (C) RISE image derived from Raman spectra and SEM data.
Monolayer

Twisted bilayer graphene

CVD-grown, bilayer graphene (here on Si/SiO2) isoften twisted and folded. These structural properties
cannot be differentiated by SEM or EDS. However,variation in peak intensities of graphene’s Raman
bands and changes in the FWHM (full width at halfmaximum) of the bands allow for the determinationof the stacking order and the twist angles of thelayers. These features can be correlated with thestructural features by RISE microscopy.

2L: AB
2L: Rotation >20°
2L: Rotation 3-8°
2L: Rotation 0-3°
2L: Rotation 12°
More than 2L

Principle of RISE Microscopy®

Samples are automatically transferred from one measuring position
to the other within the vacuum chamber of the combined Raman-SEM
instrument, streamlining the workflow and drastically improving easeof use.

The Raman Principle•

A Raman spectrum describes the energy shift of theexcitation light (laser) as a result of inelastic scatteringby the molecular bonds in a sample.

•Each molecule and chemical compound produces aparticular Raman spectrum when excited and can beeasily identified by this unique ‘fingerprint’.

•Raman spectroscopy is a well-established andnondestructive method for analyzing the molecularcomposition of a sample.

abc

Inelastic scattering of light by a molecule

dAdditional sample information from Raman spectra:
a. Peak intensity: Quantity of a specific compoundb. Peak shift: Identification of stress and strain statesc. Peak width: Degree of crystallinity
d. Polarization state: Crystal symmetry and orientation

RISE microscopy measurement ofa pearl’s surface: Raman imaging attributes
aragonite (blue, red) and vaterite (green), two polymorphs ofcalcium carbonate, to the structural features revealed by SEM.

6 µm

RISE Microscopy®

RISE systems combine all features of a stand-alone SEM and a WITec research-grade
confocal Raman imaging microscope within one instrument to provide:
Quick and convenient switching between Raman and SEM modes.
Automated sample transfer from one measuring position to the other within
the vacuum chamber.
• An integrated software interface for user-friendly measurement control.
• Easy correlation of the experimental results and image overlay.
• SEM and Raman imaging capabilities without compromise.
• A truly confocal optical path.
• Research-grade optical imaging.
••

Common vacuum chamber
Fully integrated chemical and structural
analyses accelerate correlativemeasurements.

Confocal Raman Imaging
WITec confocal Raman imaging systems combine Raman spectroscopy with confocal
microscopy to offer:
Complete Raman spectral acquisition at every image pixel with diffraction-limited
resolution (~300 nm).
• Unprecedented performance in speed, sensitivity and resolution.
• Outstanding depth resolution ideally suited to 3D image generation and depth profiles.
• Ultrahigh-throughput spectroscopic capability for the highest sensitivity.
• Nondestructive imaging. No staining or other specialized sample preparation is required.
•Scan table
The sample remains inside the vacuum
chamber during both the SEM andRaman measurements to ensure a
streamlined workflow.

Benefitsof combined RISE microscopy and energydispersive X-ray spectroscopy (EDS)®

Take full advantage of Oxford Instruments’ analytical tools for SEM systems to characterize
the chemical properties of a sample. EDS can detect elemental composition and Raman
spectroscopy can identify molecules and their bonding characteristics.

RISEEDS®

Detailed insight

Molecular and bonding information,
crystallinity, material stress and strain,
crystal orientation

Elemental quantitation and distribution,
microstructure and crystal structure

Wide-ranging analysis

Sub-micron and large-area
molecular information,
3D chemical imaging

Point analysis,large area mapping,particle analysis

Dynamic vision

Analysis of chemical processes
possible

Live Chemical Imaging in real timewith AZtecLive

Multimode operation

Independent Raman operation,
Raman-SEM image correlation

Simultaneous SEM andEDS data acquisitionA

Geoscience samplebAlFe

Intensity (a.u.)

QuartzEpidotePlagioclase (Albite)

NaSi

Raman shift (cm-1)

COxford Instruments
NanoAnalysis detectors usedin electron microscopes(A) Distribution of elements,
measured in EDS mode of the SEM.
(B) Raman spectra of the same
sample area: Quartz (brown),epidote (pink), plagioclase (green).
(C) RISE image showing thedistribution of the molecularcompounds in the sample. Raman
imaging parameters: 22,500 spectra,
80 ms integration time per spectrum

Applications
Abalone shellb(A) RISE image of a polished cross section reveals
the layered structure of the nacre (mother-of-pearl).
It consists of aragonite, a crystal form of calciumcarbonate.
(B) Raman spectra can enable the differentiationof crystal orientations (blue, red), revealing theanisotropy of the aragonite phase.

Intensity (a.u.)aRaman shift (cm-1)

BC
Intensity (a.u.)aRaman shift (cm-1)

Iron mineralogy(A) In the SEM image a piece of hematite (Fe2O3) shows some structural characteristics. (B) Hematite, goethite and vonsenite were identified from their
Raman spectra. The spectra of crystal forms of hematite are depicted in green, orange and purple, those of goethite (FeO(OH)) in red and cyan. Thespectrum of vonsenite (Fe2+2Fe3+(BO3)O2) is shown in blue. (C) Correlation of Raman and SEM data resulted in the RISE image.
abwhite brain matter
Intensity (a.u.)gray brain matter

Raman shift (cm -1)(A) RISE image of a hamster brain tissue sample. In the color-coded Raman image the whitebrain matter is shown in green and the gray matter in red. (B) The corresponding Raman
spectra reveal the different spectral characteristics of the white and gray brain matter.

BThe color-coded Raman image, showing the gold substrate(yellow) that can be clearly distinguished from GaAs (red),is correlated with the ultrastructure of the sample.

CD

Intensity (a.u.)

aLT GaAs sample

Intensity (a.u.)

Brain tissue

Raman shift (cm -1 )Cathode of a fast-cycled Li-ion battery(A) Cross section of an uncycled cathode. Particles embedded in amorphous carbon (red) contain uniformly-distributed
lithium nickel cobalt manganese oxide (Li-NMC, blue). (B) Cross section of a cycled and rapidly charged cathode.
Compared to the uncycled cathode the particles appear inhomogeneous. One particle (bottom left) shows a completely
different composition, including a spectrum (blue) identical to the Li-NMC spectrum of the uncycled electrode.
(C) Typical Raman spectra of Li-NMC (blue) and amorphous carbon (red) of the uncycled cathode. (D) Raman spectraof molecules detected in the cycled cathode. Changes in peak positions and peak width indicate variance of Li-NMCcomposition. (E) RISE image of a particle of a cycled cathode that reveals changes in Li-NMC composition andsubstantial structural degradation.
Sample courtesy of Dean Miller (Tescan USA)

Raman shift (cm -1 )Ealpha300 apyron™:
Automated Confocal
Raman Microscopealpha300 access:Confocal MicroRaman System

We take care – WITec uses environmentally friendly printed materials.
While this policy is only a small contribution to a healthy environment, we at
WITec believe that focusing on details can effect positive change in the world.

WITec Microscopesalpha300 R:
Confocal Raman
Microscopealpha300 RA:
Correlative Raman-AFM
Microscopealpha300 Ri: Inverted
Confocal Raman
Microscope

RISE®: Correlative Raman-SEM Imaging

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contact at https://raman.oxinst.com/contact
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Phone +49 (0) 731 140 700, Fax +49 (0) 731 14070 200
info@witec.de, https://raman.oxinst.com

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